LOCUS       40924_6522        7548 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBIND-GR, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7548)
  AUTHORS   Kopish K, Pagiuo A, TenHarmsel A.
  TITLE     GloResponse(TM) 9XGAL4UAS-luc2P HEK293 Cell Line Technical Bulletin,
            TB552
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7548)
  AUTHORS   Kopish K, Pagiuo A, TenHarmsel A.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-JUN-2009) Scientific Communications, Promega 
            Corporation, 5500 E. Cheryl Parkway, Madison, WI 53711, USA
REFERENCE   3  (bases 1 to 7548)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7548)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (02-JUN-2009) Scientific Communications, Promega Corporation, 5500 
            E. Cheryl Parkway, Madison, WI 53711, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7548
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        139..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1083..1523
                     /codon_start=1
                     /label=GAL4 DNA binding domain
                     /note="DNA binding domain of the GAL4 transcriptional
                     activator"
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVA"
     misc_feature    1521..1553
                     /label=IgA linker
                     /note="IgA linker"
     CDS             1563..2399
                     /codon_start=1
                     /gene="GR-LBD"
                     /product="GR LBD"
                     /label=GR-LBD
                     /note="glucocorticoid receptor ligand binding domain"
                     /protein_id="ACT53148.1"
                     /translation="MQQATTGVSQETSENPGNKTIVPATLPQLTPTLVSLLEVIEPEVL
                     YAGYDSSVPDSTWRIMTTLNMLGGRQVIAAVKWAKAIPGFRNLHLDDQMTLLQYSWMFL
                     MAFALGWRSYRQSSANLLCFAPDLIINEQRMTLPCMYDQCKHMLYVSSELHRLQVSYEE
                     YLCMKTLLLLSSVPKDGLKSQELFDEIRMTYIKELGKAIVKREGNSSQNWQRFYQLTKL
                     LDSMHEVVENLLNYCFQTFLDKTMSIEFPEMLAEIITNQIPKYSNGNIKKLLFHQK"
     gene            1563..2399
                     /gene="GR-LBD"
                     /label=GR-LBD
     misc_feature    2408..2463
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     polyA_signal    complement(2568..2689)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        2934..3291
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3342..4274
                     /codon_start=1
                     /label=hRluc
                     /note="Renilla luciferase"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             4326..5114
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="IEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPV
                     LFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSS
                     HLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQG
                     LAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIAL
                     ATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5181..5229
                     /label=poly(A) signal
                     /note="synthetic polyadenylation signal"
     promoter        5385..5489
                     /label=AmpR promoter
     CDS             5490..6347
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      complement(6508..7096)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    7212..7495
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to
                     maintain plasmids as monomers"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagccag cttgaagcaa gcctcctgaa
     1081 agatgaagct actgtcttct atcgaacaag catgcgatat ttgccgactt aaaaagctca
     1141 agtgctccaa agaaaaaccg aagtgcgcca agtgtctgaa gaacaactgg gagtgtcgct
     1201 actctcccaa aaccaaaagg tctccgctga ctagggcaca tctgacagaa gtggaatcaa
     1261 ggctagaaag actggaacag ctatttctac tgatttttcc tcgagaagac cttgacatga
     1321 ttttgaaaat ggattcttta caggatataa aagcattgtt aacaggatta tttgtacaag
     1381 ataatgtgaa taaagatgcc gtcacagata gattggcttc agtggagact gatatgcctc
     1441 taacattgag acagcataga ataagtgcga catcatcatc ggaagagagt agtaacaaag
     1501 gtcaaagaca gttgactgta gcgattccgt cgacaccacc tactccctct ccagcgatcg
     1561 ccattcagca ggccactaca ggagtctcac aagaaacctc tgaaaatcct ggtaacaaaa
     1621 caatagttcc tgcaacgtta ccacaactca cccctaccct ggtgtcactg ttggaggtta
     1681 ttgaacctga agtgttatat gcaggatatg atagctctgt tccagactca acttggagga
     1741 tcatgactac gctcaacatg ttaggagggc ggcaagtgat tgcagcagtg aaatgggcaa
     1801 aggcaatacc aggtttcagg aacttacacc tggatgacca aatgacccta ctgcagtact
     1861 cctggatgtt tcttatggca tttgctctgg ggtggagatc atatagacaa tcaagtgcaa
     1921 acctgctgtg ttttgctcct gatctgatta ttaatgagca gagaatgact ctaccctgca
     1981 tgtacgacca atgtaaacac atgctgtatg tttcctctga gttacacagg cttcaggtat
     2041 cttatgaaga gtatctctgt atgaaaacct tactgcttct ctcttcagtt cctaaggacg
     2101 gtctgaagag ccaagagcta tttgatgaaa ttagaatgac ctacatcaaa gagctaggaa
     2161 aagccattgt caagagggaa ggaaactcca gccagaactg gcagcggttt tatcaactga
     2221 caaaactctt ggattctatg catgaagtgg ttgaaaatct ccttaactat tgcttccaaa
     2281 catttttgga taagaccatg agtattgaat tccccgagat gttagctgaa atcatcacca
     2341 atcagatacc aaaatattca aatggaaata tcaaaaaact tctgtttcat caaaagtgag
     2401 tttaaacgaa ttcgggctcg gtacccgggg atcctctaga gtcgacctgc aggcatgcaa
     2461 gctgatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc caccgctgag
     2521 caataactag cataacccct tggggcggcc gcttcgagca gacatgataa gatacattga
     2581 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     2641 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     2701 ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt taagcaagta
     2761 aaacctctac aaatgtggta aaatcgaatt ttaacaaaat attaacgctt acaatttcct
     2821 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gcggatctgc
     2881 gcagcaccat ggcctgaaat aacctctgaa agaggaactt ggttaggtac cttctgaggc
     2941 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3001 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3061 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3121 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3181 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     3241 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagcttgat
     3301 tcttctgaca caacagtctc gaaccaaagg ctggagccac catggcttcc aaggtgtacg
     3361 accccgagca acgcaaacgc atgatcactg ggcctcagtg gtgggctcgc tgcaagcaaa
     3421 tgaacgtgct ggactccttc atcaactact atgattccga gaagcacgcc gagaacgccg
     3481 tgatttttct gcatggtaac gctgcctcca gctacctgtg gaggcacgtc gtgcctcaca
     3541 tcgagcccgt ggctagatgc atcatccctg atctgatcgg aatgggtaag tccggcaaga
     3601 gcgggaatgg ctcatatcgc ctcctggatc actacaagta cctcaccgct tggttcgagc
     3661 tgctgaacct tccaaagaaa atcatctttg tgggccacga ctggggggct tgtctggcct
     3721 ttcactactc ctacgagcac caagacaaga tcaaggccat cgtccatgct gagagtgtcg
     3781 tggacgtgat cgagtcctgg gacgagtggc ctgacatcga ggaggatatc gccctgatca
     3841 agagcgaaga gggcgagaaa atggtgcttg agaataactt cttcgtcgag accatgctcc
     3901 caagcaagat catgcggaaa ctggagcctg aggagttcgc tgcctacctg gagccattca
     3961 aggagaaggg cgaggttaga cggcctaccc tctcctggcc tcgcgagatc cctctcgtta
     4021 agggaggcaa gcccgacgtc gtccagattg tccgcaacta caacgcctac cttcgggcca
     4081 gcgacgatct gcctaagatg ttcatcgagt ccgaccctgg gttcttttcc aacgctattg
     4141 tcgagggagc taagaagttc cctaacaccg agttcgtgaa ggtgaagggc ctccacttca
     4201 gccaggagga cgctccagat gaaatgggta agtacatcaa gagcttcgtg gagcgcgtgc
     4261 tgaagaacga gcagaccggt ggtgggagcg gaggtggcgg atcaggtggc ggaggctccg
     4321 gagggattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4381 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4441 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4501 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4561 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4621 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4681 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4741 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4801 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     4861 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4921 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4981 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     5041 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     5101 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     5161 cctgccatca cgatggccgc aataaaatat ctttattttc attacatctg tgtgttggtt
     5221 ttttgtgtga atcgatagcg ataaggatcc tctttgcgct tgcgttttcc cttgtccaga
     5281 tagcccagta gctgacattc atccggggtc agcaccgttt ctgcggactg gctttctacg
     5341 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     5401 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     5461 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     5521 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     5581 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     5641 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttca
     5701 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     5761 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     5821 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     5881 ctgcggccaa cttacttctg acaactatcg gaggaccgaa ggagctaacc gcttttttgc
     5941 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     6001 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     6061 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     6121 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     6181 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     6241 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     6301 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ttcgaaatga
     6361 ccgaccaagc gacgcccaac cggtatcagc tcactcaaag gcggtaatac ggttatccac
     6421 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     6481 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6541 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6601 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6661 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     6721 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6781 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6841 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6901 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     6961 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     7021 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     7081 aaaaaaagga tttcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     7141 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     7201 ccttttatag tccggaaata caggaacgca cgctggatgg cccttcgctg ggatggtgaa
     7261 accatgaaaa atggcagctt cagtggatta agtgggggta atgtggcctg taccctctgg
     7321 ttgcataggt attcatacgg ttaaaattta tcaggcgcga ttgcggcagt ttttcgggtg
     7381 gtttgttgcc atttttacct gtctgctgcc gtgatcgcgc tgaacgcgtt ttagcggtgc
     7441 gtacaattaa gggattatgg taaatccact tactgtctgc cctcgtagcc atcgagataa
     7501 accgcagtac tccggccacg atgcgtccgg cgtagaggat cgagatct
//
